Role of metallothionein in nitric oxide signaling as revealed by a green fluorescent fusion protein

• DOI: 10.1073/pnas.97.1.477
• Publication Date: 2002-07-26T14:41:48Z
• Authors: Linda L. Pearce; Robin E. Gandley; Weiping Han; Karla Wasserloos; Molly Stitt; Anthony J. Kanai; Margaret K. McLau...; Bruce R. Pitt; Edwin S. Levitan
• Keywords: Metallothionein; Yellow fluorescent protein

Although the function of metallothionein (MT), a 6- to 7-kDa cysteine-rich metal binding protein, remains unclear, it has been suggested from in vitro studies that MT is an important component intracellular redox signaling, including being target for nitric oxide (NO). To directly study interaction between and NO live cells, we generated fusion protein consisting sandwiched two mutant green fluorescent proteins (GFPs). In with this chimera (FRET-MT) demonstrate resonance energy transfer (FRET) can be used follow conformational changes indicative release MT. Imaging experiments endothelial cells show agents increase cytoplasmic Ca 2+ act via endogenously rapidly persistently A role intact tissue supported by finding myogenic reflex mesenteric arteries absent knockout mice (MT −/− ) unless endogenous synthesis blocked. These results are first application intramolecular (GFP)-based FRET native utility FRET-MT as surrogate indicator production. addition, thiolate clusters signaling vascular revealed.